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Home > Substrate Dependent Microbial Biosynthesis of Auxins and Their Effect on Growth and Yield of Mung Bean Vigna Radiata L.

Substrate Dependent Microbial Biosynthesis of Auxins and Their Effect on Growth and Yield of Mung Bean Vigna Radiata L.

Thesis Info

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Author

Muhammad Ashfaq Anjum

Program

PhD

Institute

University of Agriculture

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2011

Thesis Completion Status

Completed

Subject

Applied Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/1563

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727412971

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Rhizobia are capable of promoting the growth of legumes through a number of mechanisms of action including phytohormone production. Auxin being a plant hormone is produced by rhizobia and this ability increases many folds in supplementation with some suitable physiological precursor like L-tryptophan. Substrate-derived auxins have the ability to promote plant growth by supplementing sub-optimal auxin levels in legume plants like mung bean. So, this project was designed to evaluate the effectiveness of rhizobia under axenic conditions in supplementation with L-tryptophan on the growth and yield of mung bean crop. To test this hypothesis, a series of laboratory, wire house and field experiments were conducted. Rhizobium phaseoli were isolated from different locations of Faisalabad from nodules of mung bean crop. Forty four rhizobial cultures were isolated from mung bean nodules and tested for auxins biosynthesis in vitro colorimetrically and confirmed by high performance liquid chromatography (HPLC). Rhizobial isolates varied in auxin biosynthesis ranging from 8.8 μg IAA equivalents mL -1 to 38.3 μg IAA equivalents mL -1 without L-tryptophan. Auxin biosynthesis by rhizobial isolates in supplementation with L-tyrptophan, increased many folds (1.1 to 7.9 folds) and N42 produced maximum auxins in L-tryptophan supplemented media. Different environmental factors like substrate (L-Trp) and C-source (glucose), pH, incubation temperature, incubation time and aeration (static vs. shaking) significantly affected auxin production. Growth pouch experiments were conducted in the laboratory using 34 rhizobial isolates for improving growth of mung bean seedlings using three mung bean cultivars i.e. NM-92, NM-98 and NM-2006 under axenic conditions. Fifteen isolates were selected through screening in jars using principal component scoring method. Three most efficient rhizobial isolates i. e. A23, N12 and N42 were tested alone and in combination with different L-TRP concentrations (10 -3 , 10 -4 and 10 -5 M) to test the hypothesis of substrate dependent auxin production for improving the growth of mung bean (var: NM-92, NM-98 and NM-2006) under axenic conditions. These isolates were further tested alone and in combination with 10 -3 , 10 -4 and 10 -5 M L-Trp for improving growth and nodulation of mung bean. Finally these three rhizobial isolates i.e. A23, N12 and N42 along with 10 -3 , 10 -4 , 10 -5 M L-Trp concentrations were further used in pot and field experiments using mung bean variety NM-2006. Rhizobial isolates A23, N12 and N42 alone and in combination with 10 -3 , 10 -4 and 10 -5 M L-Trp concentrations were tested in pots at the wirehouse, Institute of Soil and Environmental Sciences, University of Agriculture, Faisalabad. Results of pot experiments revealed that separate application of rhizobia significantly increased growth parameters but the combined application of rhizobia and L-tryptophan further significantly increased yield and nodulation. Field trials were conducted at two sites these rhizobial isolates A23, N12 and N42 alone and in combination with 10 -3 , 10 -4 and 10 -5 M L-Trp concentrations were tested at two N levels i. e. half and full recommended dose of N fertilizer. Results revealed that rhizobial inoculation alone significantly increased growth parameters (fresh biomass, grain yield, shoot length, root length, number of pods plant -1 , and number of grains pod -1 , 1000-grain weight, number of nodules plant -1 , fresh and dry weight of nodules, NPK contents in grain) of mung bean variety NM-2006. But the combined application of rhizobia along with L-Trp (10 -4 and 10 -5 M) further increased the mung bean yield and nodulation. This implies that substrate dependent microbial biosynthesis of auxins as a result of precursor (L-Trp)-inoculum (rhizobia) interactions can be successfully used for improving the growth and yield of legumes.
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پروفیسر سید مسعود حسن

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فتوی اور فتاوی الکاملیہ کا تعارف و اہمیت

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Bioconversion of Plant Feedstock to Fermentable Saccharides

Bioconversion of different pre-treated agricultural feed stocks into saccharides was carried out using thermophilic cellulases. Agricultural by-products i.e. sugarcane bagasse, wheat and rice straw were subjected to acid and alkali at steaming temperatures, auto-hydrolysis and microwave pre-treatments. Auto-hydrolysed and alkali pre-treated substrates showed better delignification i.e. 86.01% (wheat straw), 77.84% (sugarcane bagasse), 67.23% (rice straw) vs. 84.11% (wheat straw), 73.90% (sugarcane bagasse), 68.47% (rice straw), respectively. These de-lignified substrates were analyzed for total phenolic content (TPC) which were subsequently removed by washing with distilled water, extraction by organic solvents and detoxification by calcium hydroxide. Minimum residual TPC was observed in auto-hydrolysed substrates using calcium hydroxide i.e. 52 mg gallic acid equivalent (GAE) per g of dry weight (DW) of substrate in wheat straw, 59 mg GAE/g DW in sugarcane bagasse and 54 mg GAE/g DW in rice straw. The surface area i.e. 4 and 2mm granule size of all auto-hydrolysed substrates was assessed for saccharification potential and granule size of 2mm was found best with saccharification of 11.01% (sugarcane bagasse), 9.34% (wheat straw) and 4.36% (rice straw). Saccharification of auto-hydrolysed substrates using simultaneous addition of cellulases gave maximum saccharification in wheat straw i.e. 22.93% after 5 h of incubation at 80°C employing sodium citrate buffer of pH 6.5. Cellulase concentration of Endo-1,4-β-glucanase (125 U), Exo-1,4-β-glucanase (150 U) and β-1,4- Glucosidase (50 U) was optimized for 2% of wheat straw yielding 36.78% saccharification. Sequential addition of cellulases showed an improved saccharification of 55.64% employing Endo- 1,4-β-glucanase (75 U), Exo-1,4-β-glucanase (100 U) and β-1,4-glucosidase (50U) after 5.5 h of incubation. Scale up of the sequential saccharification in a 50 L reaction vessel resulted in increased saccharification of 57.91% with decreased time of incubation (3.5 h). In addition, substrate concentration of 2.5 % with agitation of 100 rpm was optimized with consequent saccharification of 62.12%. After saccharification, the hydrolysate was analyzed for TPC which were removed using activated charcoal. Minimum TPC i.e. 64.43 mg GAE/mL of liquid phase was achieved using 3% activated charcoal at 40°C after 20 min of incubation. The hydrolysate was analyzed by thin layer chromatography and found to have glucose and cellobiose. Wheat straw before auto-hydrolysis, after auto- hydrolysis and enzymatic breakdown was assessed for structural variance by scanning electron microscopy.